Inhibition of viral gene expression using small interfering RNA

Abstract

The invention provides methods, compositions, and kits comprising small interfering RNA (shRNA or siRNA) that are useful for inhibition of viral-mediated gene expression. Small interfering RNAs as described herein can be used in methods of treatment of HCV infection. ShRNA and siRNA constructs targetING the internal ribosome entry site (IRES) sequence of HCV are described.

We claim: 
     
       1. A method of inhibiting hepatitis C virus (HCV) RNA replication or gene expression in a cell, comprising introducing a small interfering RNA into an HCV-containing cell, wherein the antisense sequence of said small interfering RNA consists of a sequence that recognizes a sequence of 19 to 25 nucleotides within the region depicted in nucleotides 344-368 of SEQ ID NO: 11, wherein said antisense sequence is selected from the group consisting of SEQ ID NO:21, SEQ ID NO:22, SEQ ID NO: 23, and SEQ ID NO:24 and the antisense sequence of SEQ ID NO: 12, and wherein said small interfering RNA inhibits HCV RNA replication or gene expression in the cell. 
     
     
       2. A method according to  claim 1 , wherein said small interfering RNA is an shRNA. 
     
     
       3. A method according to  claim 1 , wherein said small interfering RNA is an siRNA. 
     
     
       4. A method according to  claim 1 , wherein said hepatitis C virus is genotype 1a. 
     
     
       5. A method of treating a hepatitis C virus (HCV) infection in a mammal, said method comprising administering to said mammal a composition comprising a therapeutically effective amount of a small interfering RNA wherein the antisense sequence of said small interfering RNA consists of a sequence that recognizes a sequence of 19 to 25 nucleotides within the region depicted in nucleotides 344-368 of SEQ ID NO:11, wherein said antisense sequence is selected from the group consisting of SEQ ID NO:21, SEQ ID NO:22, SEQ ID NO: 23, and SEQ ID NO:24 and the antisense sequence of SEQ ID NO: 12, and wherein said small interfering RNA inhibits HCV RNA replication or gene expression in said mammal, thereby treating HCV infection in said mammal. 
     
     
       6. A method according to  claim 1 , wherein said small interfering RNA is an shRNA. 
     
     
       7. A method according to  claim 1 , wherein said small interfering RNA is an siRNA. 
     
     
       8. A method according to  claim 1 , wherein said hepatitis C virus is genotype 1a. 
     
     
       9. A method according to  claim 1 , wherein said mammal is a human. 
     
     
       10. A composition comprising a small interfering RNA, wherein the antisense sequence of said small interfering RNA consists of a sequence that recognizes a sequence of 19 to 25 nucleotides within the region depicted in nucleotides 344-368 of SEQ ID NO:11, wherein said antisense sequence is selected from the group consisting of SEQ ID NO:21, SEQ ID NO:22, SEQ ID NO: 23, and SEQ ID NO:24 and the antisense sequence of SEQ ID NO: 12. 
     
     
       11. A composition according to  claim 10 , wherein said small interfering RNA is an shRNA. 
     
     
       12. A composition according to  claim 10 , wherein said small interfering RNA is an siRNA. 
     
     
       13. A composition according to  claim 10 , wherein said composition is a pharmaceutical composition, said pharmaceutical composition further comprising a pharmaceutically acceptable excipient. 
     
     
       14. A kit comprising a small interfering RNA, wherein the antisense sequence of said small interfering RNA consists of a sequence that recognizes a sequence of 19 to 25 nucleotides within the region depicted in nucleotides 344-368 of SEQ ID NO:11, wherein said antisense sequence is selected from the group consisting of SEQ ID NO:21, SEQ ID NO:22, SEQ ID NO: 23, and SEQ ID NO:24 and the antisense sequence of SEQ ID NO: 12. 
     
     
       15. A kit according to  claim 14 , wherein said small interfering RNA is an shRNA. 
     
     
       16. A kit according to  claim 14 , wherein said small interfering RNA is an siRNA.